Field of Invention
The present invention relates to medical technology, and more particularly related to using the raw materials for chemical testing or chemical engineering—basic fuchsine, as the key active ingredient and processing with traditional Chinese medicine and food article to manufacture a drug to cure tuberculosis and leprosy—a drug to kill acid-fast bacilli (red stain).
Description of Related Arts
Tuberculosis is a disease caused by Mycobacterium tuberculosis. Mycobacterium tuberculosis is a pathogen which causes infection inside the cell. Mycobacterium tuberculosis can adapt to acidic environment, is resistance against phagosomes of macrophages for a long period of time and can escape against the acidification of environment such that Mycobacterium tuberculosis can survive inside the body for an extended long period of time. Therefore, tuberculosis is a persistent and chronic infectious disease. Mycobacterium tuberculosis enters into the body from the respiratory tract, and through the blood stream, it spreads and bleeds in different organs, or even into the bone or lymph nodes. At present, the most common disease being caused is pulmonary tuberculosis, which is airborne and can be widely spread and impose great health hazards. Tuberculosis can be latent, carriers may have no obvious symptoms and therefore is hard to prevent. It is a big challenge and a long lasting health hazard to human.
Because of the emergence of resistant strains and the application of immunosuppression agents, the tuberculosis outbreak has worsen suddenly at worldwide level. According to WHO statistics, about one third of the world's population is infected by M. tuberculosis, and in some developing countries, the TB carriers in adults can reach 80% of which 5-8% TB carriers can develop into active TB disease.
At present, about eight million new cases of TB occur every year at worldwide level and causes the death of about three million people. In China, two hundred and fifty thousand people are died of TB every year, which is twice as the total number of death caused by transmissible diseases of all kinds. According, TB is a real worldwide health hazards to human.
M. tuberculosis enters into human body through the respiratory tract, survives and replicates inside the alveolar macrophages, then further spreads to proximal unactivated alveoli to form a Ghon focus. During the cell mediated immune response, M. Tuberculosis grows inside the macrophages and tubercles are formed of which the center of the tubercle is characterized by “caseation necrosis”, that the tubercles are capable of restricting the multiplication of M Tuberculosis. For the majority of carriers of M. tuberculosis, the M. tuberculosis and its host are in a state of mutually coexistence, that the M. tuberculosis is surrounded by fiber inside the cheesy necrotic lesion and cannot undergo multiplication. However, when the human immune system is damaged and the human body is weakened, the restricted M. tuberculosis will be re-activated and replicate, a large number of M. tuberculosis will be released from the liquefied tubercles and spread throughout the human body. These patients with primary or secondary infection require drug treatment. The conventional first line drugs includes: 1. isoniazid, 2. rifampin, 3. streptomycin 4. pyrazinamide 5. ethambutol.
All these drugs have serious toxic side effect (For example, isoniazid and rifampin can easily cause elevated serum alanine aminotransferase, hepatomegaly, jaundice, nausea, vomiting, streptomycin can cause auditory nerve damage, ethambutol can caused optic nerve damage). In additional, during the treatment process, these drugs do not kill the bacteria and allow the bacteria to remain inside the body while only inhibiting its growth. The therapeutic effect is a negative result for sputum Mycobacterium tuberculosis (cannot detect acid-fast bacilli), that the bacteria is surrounded and hidden inside the human body. Once the body immune system is weak or is infected by an influenza, the bacteria will be activated and grow again, which is a painful process. The only treatment, again, is the drug treatment. Since the drug treatment may cause toxic side effect to the patients and causes damage to liver, kidney, nervous system, gastrointestinal and hematopoietic organ, the side effect may include extremely abnormal level of aspartate aminotransferase, alanine aminotransferase, urea, creatinine, urea nitrogen and etc. In some cases, hematuria may occur and the practitioner has to switch to liver and kidney treatment from TB treatment. Many TB patients cannot recover from TB but a great damage to liver and kidney is caused.
Leprosy:
Mycobacterium leprae is the pathogen which causes leprosy. The difference between this bacteria and other mycobacteria is that this bacteria cannot be cultured in artificial culture base. Therefore, the diagnosis of leprosy can only be relied on smear examination of acid-fast stain.
The common characteristics of mycobacterium leprae and mycobacterium tuberculosis are difficult to stain and resistance to decolorization by acid and alcohol after accepting a stain, therefore they are also referred to as acid-fast bacilli. Both of them do not have flagella, spore and capsule, and are Gram (G) positive bacilli (Red stain after acid-fast staining (+)).
In taxonomy, both Mycobacterium leprae and Mycobacterium tuberculosis are classified under the class Actinobacteria, the order Actinomycetales and the branch Mycobacterium. The differences between Mycobacterium leprae and Mycobacterium tuberculosis are summarized in Table 1:
TABLE 1GramAcid-InfectionGrowthInfectionIn vitrostainFastNameMethodTemp.sitecultivationTransmission(G+)StainMycobacteriumInside37° C.InternalSuccessfulAirPositivePositivetuberculosisphagocyteorgans(Red)MycobacteriumInside28° C.BodyUn-AirPositivePositivelepraephagocyteSurfacesuccessfulContacts(Red)(Skin)
At present, the drugs for Mycobacterium leprae are basically the same as the drugs for Mycobacterium tuberculosis except that the dosage for Mycobacterium leprae is much higher and the treatment course is longer. The commonly used drugs includes rifampin, chlorophenol diazoxide, sulfamide and etc., which are highly toxic drugs and may cause serious damage to liver, kidney, nervous system, gastrointestinal and hematopoietic organ.
Basic Fuchsine (also known as: new fuchsine, fuchsine salt, rosaniline hydrochloride):
1. Basic use of basic fuchsine
Basic fuchsine is the strongest nuclear stain which can stain particles of viscoelastic tissue and fuchsinophilic substances, and is a nuclear stain for central nervous system. In bacteriology, it is used to identify Mycobacterium tuberculosis. In analytical chemistry, it is used to prepare Schiff's reagent for aldehydes testing. In bromate titration, it is used as an redox indicator. It is also used for dyeing cotton, linen, artificial fibers, leather, feathers, fat and etc. as well as for manufacture of color precipitates.
2. Sources of Preparation
In the presence of iron and zinc chloride, react hydrochlorides of aniline, o-toluidine and p-toluidine with nitrobenzene for condensation, then separate by acid-base extraction and obtain by crystallization.
3. Chemical structure

Description of Structure:
For use as a stain for bacteria, the majority of synthetic substances are organic compounds containing a benzene ring (Basic fuchsine is one of the kind) in which a chromophore group and an auxochrome group are provided on the benzene ring. The chromophore group includes a nitro group (—NO2) and an azo group (—N═N—), while the auxochrome group includes a hydroxyl (—OH) and an amino group (—NH2). The chromophore group is responsible for the color of a compound and a benzene-containing compound with a chromophore group is called a chromogen. This type of compounds is not a dye because it does not have any affinity to the dying object and is easily separated from the dying object. Therefore, the auxochrome group is required. The auxochrome group does not contribute any color to the dyeing object but has the ability of dissociation to provide ionic character to a compound. After dissociation, the dye materials can bind to the dyeing object and provide color to the dyeing object.
The auxochrome group may be a basic group (such as a —NH2 group) or an acid group (such as a —OH group). The more the —NH2 group or the —OH group, the stronger is the basicity or acidity. If the number of each group is one, then the basic group will dominate. Accordingly, the pH of the dye can be determined by the nature of the auxochrome group. The basic fuchsine has no —OH group in its structure, and therefore is a basic dye.
Because organic dye is an organic acid or base with color which is difficult to dissolve in water and is soluble in organic solvent, usually they are made into salt in order to make them easier to dissolve in water. Therefore, basic fuchsine is also called fuchsine salt. In conclusion, the only difference between the above two formulae is just one auxochrome group (CH2), which is a substance existed in the form of two structures in solution.
Principle of Acid-Fast Bacilli Stain
Acid-fast bacilli contains mycolic acid. Basic Fuchsine (C20H20N3Cl or C19H18N3Cl) has a higher solubility in lipids than in hydrochloric acid and alcohol. Therefore, both can be combined strongly and is not easily decolorized by hydrochloric acid or alcohol. In addition, maintaining the integrity of the surface cell structure during the dyeing process is an important factor for maintaining its acid resistance. It is because a complete cell wall can prevent the overflow of lipids which is stained by basic fuchsine. Therefore, the red stain of the acid-fast bacilli can be maintained even after bleaching. Non acid-fast bacilli does not contain mycolic acid, cannot resist decoloring by hydrochloric acid or alcohol and therefore can be re-stained by methylene blue into blue color.
At present, the mechanism of bacteria stain is not completely clear. According to the available evidence, the dyeing process is neither purely physical nor purely chemical, but rather a combination of both.
According to the characteristics of acid-fast bacilli, acid-fast staining is mainly used in testing of Mycobacterium tuberculosis and Mycobacterium leprae. 
4. Molecular formula and molecular weightC20H20N3CL=337.84  Formula I:C19H18N3CL=323.82  Formula II:
5. Properties: green metallic luster crystal, soluble in ethanol and amyl alcohol, slightly soluble in water, red in color in aqueous solution, insoluble in ether.